HeLa cells

I recently thawed HeLa cells and have started growing these in DMEM media. I passaged it thrice till now (once in 2days). I see some healthy but lot of round cells. I am not sure if the stock is bad or am I doing something wrong. One thing I remember is that I pipetted the cells vigourously to break the lumps during subculturing them. Can someone suggest how can I bring these cells to a healthy condition? Should I be patient and continue this for another week and see (since usually they say it takes 2 weeks to get good cells) or should I thaw another fresh stock.

I recently thawed HeLa cells and have started growing these in DMEM media. I passaged it thrice till now (once in 2days). I see some healthy but lot of round cells. I am not sure if the stock is bad or am I doing something wrong. One thing I remember is that I pipetted the cells vigourously to break the lumps during subculturing them. Can someone suggest how can I bring these cells to a healthy condition? Should I be patient and continue this for another week and see (since usually they say it takes 2 weeks to get good cells) or should I thaw another fresh stock.

Thanks in advance.

Mamcy

Are these cells that you bought or that were previously frozen in your lab? If they were frozen in-house, were they frozen in DMSO to prevent crystals? Depending on how much you have, you could thaw another batch, but if you still have some viable cells, I'd recommend that you give them a bit longer to recover from being frozen. Also, how were the cells stored while frozen?

Good luck

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then heaven will be yours, before you meet your end

If the cells are not doing well, it is best to start again, proceeding in an experiment with cells that are sick or dying may well affect the results you get. Continuing to grow these cells can also result in laboratory selection for a more resistant/tougher cell population, that have diverged from the parent line, meaning that your experimental results may be different from results from another lab using the same techniques and supposedly the same cells.

Hi, don't worry about! I often have the same problem with HeLa cells. You simply need to spread cells pipetting a small number of cells for each flask/dish and be patient. When you use trypsin, you have to leave flasks/dishes for 2 min at 37°C to avoid mechanical aid in detachment. All will be ok in a week! Good luck!!!